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The remarkable ability to target one or even a couple of sites on a
large protein with a small molecule, under photochemical control,
is a considerable challenge and this challenge has been addressed
in some depth in this book. Systematic modification of the
structure of the photoreagent provided valuable information on the
binding site recognition as well as the mechanism of the
photocleavage reaction. Some factors that impact the photocleavage
reaction include the exact location of the probe binding site on
the protein, conformations of the bound probe at the binding site,
protein size, functional groups present on the probe that interact
with the protein surroundings either in a favorable or unfavorable
manner, overall charge on the photoreagent, and photophysical as
well as photochemical properties of the probe. The protein
photocleavage studies, in all case, were preceded by detailed
binding studies by a variety of spectroscopic methods. Methods as
simple as absorption and fluorescence spectroscopies or more
sophisticated circular dichroism spectroscopy were used.
Conclusions that are most consistent with the binding data
indicated a single binding site on most proteins, irrespective of
the probe or the protein, with only one exception noted so far.
Photoactivation of the bound probe resulted in protein cleavage at
a single site, in many instances. The specificity for the reaction
has been investigated in detail and molecular modeling studies
provided a firm ground to rationalize the observed cleavage sites.
The reagents provide unique tools for sequencing large proteins by
converting them into smaller fragments by non-biochemical
transformations. Understanding of the rules for the above
methodology are also investigated which provided rational methods
for the design of small molecules that could bind at particular
sites on large proteins, and this is a major breakthrough for a
variety of fields including drug design, protein targeting, mass
spectrometry, proteomics and other cutting-edge research areas.
This is the second volume in the series of books covering practical
aspects of synthesis and characterization of various categories of
nanomaterials taking into consideration the most up to date
research publications. The aim of the book series is to provide
students and researchers practical information such as synthetic
procedures, characterization protocols and mechanistic insights to
enable them to either reproduce well established methods or plan
for new syntheses of size and shaped controlled nanomaterials. The
second volume focuses on multifunctional nanomaterials.
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